An Endogenously Tagged Fluorescent Fusion Protein Library in Mouse Embryonic Stem Cells.

ABSTRACT: Embryonic stem cells (ESCs), with their dual capacity to self-renew and differentiate, are commonly used to study differentiation, epigenetic regulation, lineage choices, and more. Using non-directed retroviral integration of a YFP/Cherry exon into mouse ESCs, we generated a library of over 200 endogenously tagged fluorescent fusion proteins and present several proof-of-concept applications of this library. We show the utility of this library to track proteins in living cells; screen for pluripotency-related factors; identify heterogeneously expressing proteins; measure the dynamics of endogenously labeled proteins; track proteins recruited to sites of DNA damage; pull down tagged fluorescent fusion proteins using anti-Cherry antibodies; and test for interaction partners. Thus, this librar

SUBMITTER: Harikumar A 

PROVIDER: S-EPMC5639459 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature